As an important endogenous gasotransmitter, hydrogen sulfide (H2S) plays a critical role in various physiological functions, and has been regarded as a biomarker of cancer due to its overexpression in cancer cells. In addition, the early stages of cancer are often accompanied by abnormalities in the intracellular microenvironments. However, deep insights into the simultaneous detection of H2S and viscosity/polarity variations in cancer cells/tissues are rarely reported. In this work, we developed a near-infrared (NIR) and mitochondria-targeting fluorescent probe PDQHS, which exhibits high selectivity for H2S with an emission peak around 632 nm, and excellent response to viscosity/polarity through NIR emission beyond 706 nm. Meanwhile, PDQHS shows good biocompatibility, and can specifically accumulate into mitochondria. Using PDQHS, the visual distinguishing cancer cells from normal cells was achieved via dual-channel detection of H2S and viscosity/polarity. More importantly, PDQHS has been successfully applied to visual diagnosis of tumor tissue. Obviously, this simultaneous monitoring multi-markers via dual-channel is conducive to amplify the detection signal compared to the de-tection of a single biomarker, thus providing a more sensitive and reliable imaging tool for cancer diagnosis.